SAPK4 CRISPR Activation Plasmid (h): sc-400423-ACT

MAPK13 encodes stress-activated protein kinase 4 (SAPK4/p38δ), a member of the p38 MAPK family that integrates inflammatory cytokines, oxidative stress, and environmental cues into context-dependent transcriptional and post-translational responses. SAPK4 participates in MAPK signaling networks that regulate cell differentiation, apoptosis, cytoskeletal dynamics, and innate immune effector programs through phosphorylation of downstream substrates and modulation of transcription factor activity. Dysregulated MAPK13 signaling has been reported across multiple disease-relevant contexts, including inflammatory disorders and oncogenic phenotypes, where p38δ activity can influence epithelial biology and immune–stromal interactions. As a node within stress response pathways, SAPK4 is frequently studied to dissect stimulus-specific signaling crosstalk and downstream gene expression programs.

SAPK4 CRISPR Activation Plasmid (h) provides a targeted, non-destructive approach to upregulating endogenous MAPK13 expression without altering the underlying DNA sequence.

SAPK4 CRISPR Activation Plasmid (h) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the MAPK13 locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the MAPK13 transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous SAPK4 expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native MAPK13 locus and enabling the study of SAPK4-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of SAPK4 pathway restoration in tumor cells with silenced or reduced MAPK13 expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.